Histone H3 (K4 methylation) Panel (mono methyl K4, di methyl K4, tri methyl K4)

• ChIP

Unknown

ChIP - Histone H3 (K4 methylation) Panel (mono methyl K4, di methyl K4, tri methyl K4) (AB103938)

Chromatin was prepared from U2OS cells according to the Abcam X-ChIP protocol. Cells were fixed with formaldehyde for 10 min. The ChIP was performed with 25 µg of chromatin, 2 µg of ab1791 (blue), and 20 µl of protein A/G sepharose beads. No antibody was added to the beads control (yellow). The immunoprecipitated DNA was quantified by real time PCR (Taqman approach). Primers and probes are located in the first kb of the transcribed region.

• ChIP

Unknown

ChIP - Histone H3 (K4 methylation) Panel (mono methyl K4, di methyl K4, tri methyl K4) (AB103938)

Chromatin was prepared from U2OS cells according to the Abcam X-ChIP protocol. Cells were fixed with formaldehyde for 10min. The ChIP was performed with 25µg of chromatin, 2µg of ab8895 (blue), and 20µl of Protein A/G sepharose beads. No antibody was added to the beads control (yellow). The immunoprecipitated DNA was quantified on the GAPDH and ALDOA (active) and MYO-D (inactive) promoters and over the y-Actin gene (active). Schematic diagram of the y-Actin gene is shown on the top of the figure. Black boxes represent exons and thin lines represent introns. PCR products are depicted as bars under the gene.

• ChIP

Unknown

ChIP - Histone H3 (K4 methylation) Panel (mono methyl K4, di methyl K4, tri methyl K4) (AB103938)

Chromatin was prepared from Hela cells according to the Abcam X-ChIP protocol. Cells were fixed with formaldehyde for 10min. The ChIP was performed with 25µg of chromatin, 8µl of ab32356 (blue), and 20µl of Protein A/G sepharose beads. No antibody was added to the beads control (yellow). The immunoprecipitated DNA was quantified by real time PCR (Taqman approach). Primers and probes are located in the first kb of the transcribed region.

• ChIP

Unknown

ChIP - Histone H3 (K4 methylation) Panel (mono methyl K4, di methyl K4, tri methyl K4) (AB103938)

Chromatin was prepared from U2OS cells according to the Abcam X-ChIP protocol. Cells were fixed with formaldehyde for 10min. The ChIP was performed with 25 µg of chromatin, 2 µg of ab12209 (blue), and 20 µl of Protein A/G sepharose beads. No antibody was added to the beads control (yellow). The immunoprecipitated DNA was quantified by real time PCR (Taqman approach). Primers and probes are located in the first kb of the transcribed region.

• WB

Unknown

Western blot - Histone H3 (K4 methylation) Panel (mono methyl K4, di methyl K4, tri methyl K4) (AB103938)

ab8895 is specific for mono-methylated Lysine 4 of histone H3 and does not recognize di- or tri-methyl Lysine 4 nor methylation at Lysine 9. This is shown in lane 2 where the activity of the antibody is specifically blocked by the addition of the immunizing peptide (ab1340).

All lanes:

Western blot - Anti-Histone H3 (mono methyl K4) antibody - ChIP Grade (<a href='/en-us/products/primary-antibodies/histone-h3-mono-methyl-k4-antibody-chip-grade-ab8895'>ab8895</a>) at 1/500 dilution

Lane 1:

Calf thymus histone lysate

Lane 2:

Calf thymus histone lysate with Human Histone H3 (mono methyl K4) peptide (<a href='/en-us/products/proteins-peptides/human-histone-h3-mono-methyl-k4-peptide-ab1340'>ab1340</a>)

Lane 3:

Calf thymus histone lysate with Human Histone H3 (di methyl K4) peptide (<a href='/en-us/products/proteins-peptides/human-histone-h3-di-methyl-k4-peptide-ab7768'>ab7768</a>)

Lane 4:

Calf thymus histone lysate with Human Histone H3 (tri methyl K4) peptide (<a href='/en-us/products/proteins-peptides/human-histone-h3-tri-methyl-k4-peptide-ab1342'>ab1342</a>)

Lane 5:

Calf thymus histone lysate with Human Histone H3 (mono methyl K9) peptide (<a href='/en-us/products/proteins-peptides/human-histone-h3-mono-methyl-k9-peptide-ab1771'>ab1771</a>)

Lane 6:

Calf thymus histone lysate with Human Histone H3 (mono methyl K27) peptide (<a href='/en-us/products/proteins-peptides/human-histone-h3-mono-methyl-k27-peptide-ab1780'>ab1780</a>)

Lane 7:

Calf thymus histone lysate with Human Histone H3 (unmodified ) peptide (<a href='/en-us/products/proteins-peptides/human-histone-h3-unmodified-peptide-ab2903'>ab2903</a>)

Secondary

All lanes:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (<a href='/en-us/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab6721'>ab6721</a>) at 1/5000 dilution

Predicted band size: 15 kDa

Observed band size: 18 kDa

false

Exposure time: 2min